Fig 1: (a) DPV measurements of the PSA antigen in 0.1 M PBS. (b) DPV measurements of the PSA antigen in undiluted human serum. (c) Calibration curve based on the DPV measurement of the PSA antigen in undiluted human serum. (d) Interference test on the PSA biosensor using the AMACR antigen.
Fig 2: Effect of androgen treatment on PSA secretion from LNCaP cells. (A) 2000 LNCaP cells were seeded without any stimulus for 24 h. (B) 2000 cells LNCaP cells stimulated with R1881 [0.5 nM] were seeded, and secretion was captured 24 h after stimulation. (C) Overnight stimulated LNCaP cells with R1881 [0.5 nM] were treated with Enzalutamide [2 µM], and secretion was captured 24 h after inhibition. (D) 2000 LNCaP cells were seeded without any stimulus for 48 h. (E) 2000 cells LNCaP cells stimulated with R1881 [0.5 nM] were seeded, and secretion was captured 48 h after stimulation. (F) Overnight stimulated LNCaP cells with R1881 [0.5 nM] were treated with Enzalutamide [2 µM] and secretion was captured 48 h after inhibition. (G) Scatterplot showing relative PSA secretion (in arbitrary units) from single LNCaP cells after 24 and 48 h (N = 3). * p < 0.05, **** p < 0.0001, according to Kruskal–Wallis and Dunn’s post hoc test.
Fig 3: PSA secretion from LNCaP cells sorted into G1, S, and G2/M phase on PVDF membranes. (A) Highest PSA spots observed from cells in the G1 phase, (B) few secretion spots seen in S phase and (C) no secretion was observed in G2/M phase. The second row indicates magnified images of an area on the membranes presented in the first row.
Fig 4: Immunofluorescence staining of LNCaP cells. The control cells are stained with DAPI for nuclear visualization in blue (A), PSA (FITC) in green (B), and PSMA(PE) in red (C). The R1881-stimulated cells are stained similarly with DAPI (E), PSA (F), and PSMA (G). The enzalutamide-inhibited cells are stained with DAPI (I), PSA (J), and PSMA (K). The overlay of all channels for the control, R1881-stimulated and enzalutamide-inhibited cells are shown in (D), (H) and (L) respectively. Scale bar = 50 μm.
Fig 5: Effect of androgen treatment on PSA secreting LNCaPs. (A) Percentage of PSA-secreting LNCaP cells after 24 h and 48 h of androgen stimulation (blue) and inhibition (green) (N = 3). (B) Cumulative PSA secreted by LNCaP cells upon androgen activation and inhibition after 24 h and 48 h (N = 3).
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